Syntaphilin CRISPR Activation Plasmid (h): sc-403784-ACT

Human SNPH encodes syntaphilin, a mitochondria-anchoring protein that immobilizes axonal mitochondria by coupling them to the microtubule cytoskeleton. By restricting mitochondrial motility, syntaphilin helps tune local ATP availability and Ca²⁺ buffering at presynaptic sites, linking mitochondrial dynamics to synaptic maintenance and activity-dependent plasticity. SNPH function intersects with pathways governing axonal transport, mitochondrial quality control, and neuronal stress responses, making it relevant to studies of neurodegeneration and other disorders where mitochondrial distribution and bioenergetic homeostasis are perturbed. Altered regulation of mitochondrial trafficking is frequently examined in models of neuronal injury and proteinopathy, positioning SNPH as a mechanistic node for investigating circuit vulnerability.

Syntaphilin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SNPH expression without altering the underlying DNA sequence.

Syntaphilin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SNPH locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the SNPH transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Syntaphilin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SNPH locus and enabling the study of Syntaphilin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Syntaphilin pathway restoration in tumor cells with silenced or reduced SNPH expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.